title: Conditional knockout of myocyte focal adhesion kinase abrogates ischemic preconditioning in adult murine hearts.
abstract: BACKGROUND
  Our laboratory has previously demonstrated the importance of a cytoskeletal-based survival signaling pathway using in vitro models of ischemia/reperfusion (IR). However, the importance of this pathway in mediating stress-elicited survival signaling in vivo is unknown.
  
  
  RESULTS
  The essential cytoskeletal signaling pathway member focal adhesion kinase (FAK) was selectively deleted in adult cardiac myocytes using a tamoxifen-inducible Cre-Lox system (α-MHC-MerCreMer). Polymerase chain reaction (PCR) and Western blot were performed to confirm FAK knockout (KO). All mice were subjected to a 40-minute coronary occlusion followed by 24 hours of reperfusion. Ischemic preconditioning (IP) was performed using a standard protocol. Control groups included wild-type (WT) and tamoxifen-treated α-MHC-MerCreMer+/-/FAK(WT/WT) (experimental control) mice. Infarct size was expressed as a percentage of the risk region. In WT mice IP significantly enhanced the expression of activated/phosphorylated FAK by 36.3% compared to WT mice subjected to a sham experimental protocol (P ≤ 0.05; n = 6 hearts [sham], n = 4 hearts [IP]). IP significantly reduced infarct size in both WT and experimental control mice (43.7% versus 19.8%; P ≤ 0.001; 44.7% versus 17.5%; P ≤ 0.001, respectively). No difference in infarct size was observed between preconditioned FAK KO and nonpreconditioned controls (37.1% versus 43.7% versus 44.7%; FAK KO versus WT versus experimental control; P=NS). IP elicited a 67.2%/88.8% increase in activated phosphatidylinositol-3-kinase (PI3K) p85/activated Akt expression in WT mice, but failed to enhance the expression of either in preconditioned FAK KO mice.
  
  
  CONCLUSIONS
  Our results indicate that FAK is an essential mediator of IP-elicited cardioprotection and provide further support for the hypothesis that cytoskeletal-based signaling is an important component of stress-elicited survival signaling.
authors.affiliation: Department of Pathology, Louisiana State University Health Sciences Center, New Orleans, LA.
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Perricone
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Bivona
authors.forename: Fannie R
authors.initials: FR
authors.lastname: Jackson
authors.forename: Richard S
authors.initials: RS
authors.lastname: Vander Heide
mesh_terms: D000367:Age Factors; D000818:Animals; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000235:genetics / Q000502:physiology*; D019157:Ischemic Preconditioning, Myocardial; D008297:Male; D051379:Mice; D032342:Muscle Cells / Q000502:physiology*; D009206:Myocardium / Q000201:enzymology*
chemical_list: D051416:Focal Adhesion Protein-Tyrosine Kinases
keywords: cytoskeleton; ischemia; ischemic preconditioning